Preparation and in vitro characterization of carrier erythrocytes for vaccine delivery

Erythrocytes as the most readily available and abundant cells within the body, have been studied extensively for their potential application as drug delivery carriers. In this study, human erythrocytes have been loaded by bovine serum albumin (BSA) as a model antigen/protein using hypotonic preswelling method for targeted delivery of this antigen to antigen-presenting cells (APCs). A series of in vitro tests have been carried out to characterize the carrier cells in vitro, including loading parameters, BSA and hemoglobin release kinetics, hematological indices, particle size distribution, SEM analysis, osmotic and turbulence fragility, and osmotic competency. BSA was loaded in erythrocytes with a loaded amount of 1.98 ± 0.009 mg with antigen release from carrier cells showing a zero-order kinetic consistent to that of the cell lysis. The apparent cell sizes, measured using laser scattering, were not significantly different from normal erythrocytes, but the real sizes, measured using SEM, and surface topologies were quite different between loaded and unloaded cells. The BSA-loaded cells were remarkably more fragile and less deformable compared to the normal cells. Totally, BSA-loaded erythrocytes seem to be a promising delivery system for reticuloendothelial system (RES) targeting of the antigens.